title: Enhancement of enteric adenovirus cultivation in a novel Ras-overexpressing cell line.
abstract: Enteric human adenoviruses (HAdVs; serotypes 40 and 41) have been identified as an emerging cause of drinking water contamination. Due to their fastidious characteristics, HAdVs are difficult to cultivate and therefore not detected easily by standard mammalian cell cultivation methods. Here we found that human embryonic kidney 293 cells, transformed transiently with Ras, enhanced HAdV replication by more than threefold. We also constructed a stable cell line overexpressing the Ras protein, 293-Ras, in which the replication of three HAdV strains of serotypes 40 and 41 was increased markedly. However, only HAdV replication was enhanced; infection of 293 and 293-Ras cells with human rhinivorus-6 showed no significant differences in replication rate. Infected 293-Ras cells exhibited an increased level and phosphorylation of extracellular regulated kinase (ERK). In addition, the Ras-mediated increase in HAdV replication was impaired by the mitogen-activated protein kinase/ERK kinase (MEK1) inhibitor U0126, suggesting direct involvement of the MEK1/ERK pathway in enhanced HAdV replication. Based on these results, we suggest that the 293-Ras cell line be used for the efficient detection and cultivation of HAdV strains in both clinical and environmental specimens.
authors.affiliation: Department of Environmental Health, School of Public Health, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
authors.forename: Jiyeon
authors.initials: J
authors.lastname: Si
authors.affiliation: Department of Environmental Health, School of Public Health, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
authors.forename: Misoon
authors.initials: M
authors.lastname: Kim
authors.affiliation: Department of Environmental Health, School of Public Health, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
authors.forename: Mi Young
authors.initials: MY
authors.lastname: Lim
authors.affiliation: Department of Environmental Health, School of Public Health, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
authors.forename: GwangPyo
authors.initials: G
authors.lastname: Ko
mesh_terms: D000258:Adenovirus Infections, Human / Q000201:enzymology / Q000235:genetics / Q000821:virology*; D000260:Adenoviruses, Human / Q000235:genetics / Q000502:physiology*; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D006801:Humans; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism; D015689:Oncogene Protein p21(ras) / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D014776:Virus Cultivation; D014779:Virus Replication
chemical_list: D020929:Mitogen-Activated Protein Kinase Kinases; D015689:Oncogene Protein p21(ras)